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Brief Report

Single-Round Infectious Particle Production by DNA-Launched Infectious Clones of Bungowannah Pestivirus

1
Institute of Diagnostic Virology, Friedrich-Loeffler-Institut, 17493 Greifswald-Insel Riems, Germany
2
Molecular Virology Laboratory, Department of Medical Microbiology, Leiden University Medical Center, 2333 ZA Leiden, The Netherlands
*
Author to whom correspondence should be addressed.
Viruses 2020, 12(8), 847; https://doi.org/10.3390/v12080847
Received: 15 July 2020 / Revised: 31 July 2020 / Accepted: 31 July 2020 / Published: 4 August 2020
(This article belongs to the Special Issue Bovine Viral Diarrhea Virus and Related Pestiviruses)
Reverse genetics systems are powerful tools for functional studies of viral genes or for vaccine development. Here, we established DNA-launched reverse genetics for the pestivirus Bungowannah virus (BuPV), where cDNA flanked by a hammerhead ribozyme sequence at the 5′ end and the hepatitis delta ribozyme at the 3′ end was placed under the control of the CMV RNA polymerase II promoter. Infectious recombinant BuPV could be rescued from pBuPV-DNA-transfected SK-6 cells and it had very similar growth characteristics to BuPV generated by conventional RNA-based reverse genetics and wild type BuPV. Subsequently, DNA-based ERNS deleted BuPV split genomes (pBuPV∆ERNS/ERNS)—co-expressing the ERNS protein from a separate synthetic CAG promoter—were constructed and characterized in vitro. Overall, DNA-launched BuPV genomes enable a rapid and cost-effective generation of recombinant BuPV and virus mutants, however, the protein expression efficiency of the DNA-launched systems after transfection is very low and needs further optimization in the future to allow the use e.g., as vaccine platform. View Full-Text
Keywords: Bungowannah virus; flavivirus; reverse genetics; single round infectious particle Bungowannah virus; flavivirus; reverse genetics; single round infectious particle
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MDPI and ACS Style

Dalmann, A.; Wernike, K.; Snijder, E.J.; Oreshkova, N.; Reimann, I.; Beer, M. Single-Round Infectious Particle Production by DNA-Launched Infectious Clones of Bungowannah Pestivirus. Viruses 2020, 12, 847. https://doi.org/10.3390/v12080847

AMA Style

Dalmann A, Wernike K, Snijder EJ, Oreshkova N, Reimann I, Beer M. Single-Round Infectious Particle Production by DNA-Launched Infectious Clones of Bungowannah Pestivirus. Viruses. 2020; 12(8):847. https://doi.org/10.3390/v12080847

Chicago/Turabian Style

Dalmann, Anja, Kerstin Wernike, Eric J. Snijder, Nadia Oreshkova, Ilona Reimann, and Martin Beer. 2020. "Single-Round Infectious Particle Production by DNA-Launched Infectious Clones of Bungowannah Pestivirus" Viruses 12, no. 8: 847. https://doi.org/10.3390/v12080847

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